ABSTRACT
Aim: To predict glycosylphosphatidylinositol (GPI)-anchored proteins in the genome of Paracoccidioides brasiliensis and Paracoccidioides lutzii. Materials & methods: Five different bioinformatics tools were used for predicting GPI-anchored proteins; we considered as GPI-anchored proteins those detected by at least two in silico analysis methods. We also performed the proteomic analysis of P. brasiliensis cell wall by mass spectrometry. Results: Hundred GPI-anchored proteins were predicted in P. brasiliensis and P. lutzii genomes. A series of 57 proteins were classified in functional categories and 43 conserved proteins were reported with unknown functions. Four proteins identified by in silico analyses were also identified in the cell wall proteome. Conclusion: The data obtained in this study are important resources for future research of GPI-anchored proteins in Paracoccidioides spp. to identify targets for new diagnostic tools, drugs and immunological tests.
Subject(s)
Fungal Proteins/genetics , Glycosylphosphatidylinositols/metabolism , Membrane Proteins/genetics , Paracoccidioides/metabolism , Amino Acid Sequence , Cell Wall/genetics , Cell Wall/metabolism , Computational Biology , Conserved Sequence , Fungal Proteins/chemistry , Fungal Proteins/metabolism , Genome, Fungal/genetics , Membrane Proteins/chemistry , Membrane Proteins/metabolism , Open Reading Frames , Paracoccidioides/genetics , Paracoccidioides/pathogenicity , Paracoccidioidomycosis/microbiology , Proteomics , VirulenceABSTRACT
AIM: To perform the proteomic profile of Paracoccidioides lutzii after treatment with the compound camphene thiosemicarbazide (TSC-C) in order to study its mode of action. METHODS: Proteomic analysis was carried out after cells were incubated with TSC-C in a subinhibitory concentration. Validation of the proteomic results comprised the azocasein assay, western blot and determination of the susceptibility of a mutant to the compound. RESULTS: Proteins related to metabolism, energy and protein fate were regulated after treatment. In addition, TSC-C reduces the proteolytic activity of the protein extract similarly to different types of protease inhibitors. CONCLUSION: TSC-C showed encouraging antifungal activity, working as a protease inhibitor and downregulating important pathways impairing the ability of the fungi cells to produce important precursors.
